Gliomas are the most common primary tumors of the central nervous system. Among them, glioblastoma (GBM), a World Health Organization (WHO) grade 4 glioma, is the most aggressive form and remains one of the most lethal human cancers. Its poor prognosis is largely attributable to rapid progression, frequent recurrence, and profound therapeutic resistance, all of which are closely associated with glioma stem cells (GSCs). Although several signaling pathways sustaining GSC properties have been identified, upstream regulators that coordinate these pathways remain incompletely understood. TIMELESS, originally identified as a component of the circadian clock, has recently been implicated in the regulation of brain function and is aberrantly overexpressed in multiple cancer types; however, its role in glioma malignancy has not been defined. Here, we demonstrate that TIMELESS mRNA expression increases with glioma grade and is associated with poor prognosis across multiple glioma cohorts. Genetic depletion of Timeless suppressed tumor aggressiveness and prolonged survival in an orthotopic mouse glioma model by attenuating GSC properties. Mechanistically, Timeless enhanced stemness of glioma by upregulating Janus kinases (JAK) expression and promoting phosphorylation of signal transducer and activator of transcription 3 (STAT3). These effects were conserved in human GBM cells, supporting the relevance of TIMELESS-mediated signaling across species. Together, our findings uncover an unexpected, clock-independent function of TIMELESS in sustaining glioma stemness and malignancy and highlight the TIMELESS-JAK-STAT3 axis as a non-canonical mechanism that operates beyond the traditional circadian clockwork in treatment-resistant glioma.

Agonists of the lipid-sensing PPAR nuclear receptors, including PPARα, are being explored as therapies for metabolic disorders due to their roles in metabolic and anti-inflammatory processes. PPARα transcriptional programs have tissue-dependent features, yet there is a lack of genetic tools to study tissue-specific signaling activation without the addition of a systemic agonist. We aimed to investigate intestinal epithelial cell (IEC)-specific roles of PPARα signaling using a novel transgenic mouse that enables spatial and temporal control of Ppara overexpression. CAG-Ppara, -EGFP mice were bred to Villin-CreERT2 to establish the IEC-Ppara mouse, which was compared to littermate controls 2 weeks after tamoxifen exposure. IEC-Ppara mice had increased Ppara mRNA and PPARα protein in the intestinal epithelium. Transcriptional analysis of intestinal tissue from IEC-Ppara mice showed upregulation of PPARα target genes and functional enrichment for fatty acid catabolic processes. As expected, the enterocytes of IEC-Ppara mice were primed to absorb lipids following oral administration of an olive oil bolus. Unexpectedly, the enteroendocrine hormone Gip was among the most downregulated genes. GIP-positive cells were reduced in the intestines of IEC-Ppara mice and in mice treated with PPARα agonist WY-14643. Circulating GIP hormone, but not GLP-1 hormone, was reduced in IEC-Ppara mice. GLP-1-positive cells and hormone were unchanged. Consistent with reduced GIP function, IEC-Ppara mice consumed more food. These findings reveal PPARα as a regulator of GIP and support a new framework in which PPARα signaling influences systemic energy balance via a gut hormone axis. This study could have future impact on understanding responses to therapies targeting PPAR or incretin signaling.

Neural organoids provide tractable models of human brain development, function, and disease, but adeno-associated viral vector (AAV)-based gene delivery is limited by inefficient transduction and insufficient comparative evaluation of the natural and engineered capsids. Here, we evaluated seven AAV capsids (AAV-2, -6, -9, -DJ, -2-retro, -PHP.eB, and -Cap-Mac) for transduction efficiency, cell type tropism, cytotoxicity, and functional integrity in human cortical and thalamic organoids. AAV-Cap-Mac demonstrated the highest transduction efficiency, with broad cellular tropism, but caused cytotoxicity and decreased neuronal activity. By contrast, AAV-DJ yielded efficient neuronal transduction, with preserved calcium dynamics and minimal cytotoxicity, which enabled rabies virus-based circuit tracing and all-optical physiology. Our findings reveal a capsid-dependent trade-off between transduction efficiency and functional integrity and identify AAV-DJ as an optimal serotype for robust gene delivery and physiological interrogation in organoid-based neuroscience. Combining AAV-DJ with emerging molecular and circuit-level technologies in organoids should facilitate mechanistic and translational studies of the human brain.

Natural Killer (NK) cells generally exhibit dysfunction in tumor microenvironment (TME), significantly limiting their efficacy in antitumor therapy. Tumor cells display enhanced glycolysis, leading to lactic acid (LA) secretion and accumulation in the TME. Using human-induced pluripotent stem cell (hiPSC)-derived NK cells (iNKs) as a model, we demonstrate that LA induces substantial iNK dysfunction, including reduced survival, impaired IFN-γ secretion, and diminished tumor-killing capacity due to severely compromised mitochondrial function. To overcome LA-induced dysfunction, we knocked in an expression cassette for lactate dehydrogenase B (LDHB) into hiPSCs (LDHB-hiPSCs), enabling conversion of cellular lactate to pyruvate. iNKs derived from LDHB-hiPSCs (LDHB-iNKs) largely resisted LA-induced dysfunction, exhibiting enhanced cytotoxicity and improved survival in high-LA tumor tissues. Importantly, LDHB-iNKs show superior suppression of solid tumor formation in vivo. Our study provides a novel strategy to enhance NK cell therapy against solid tumors by reshaping the metabolic pathways.

Smoking cessation decreases lung cancer progression; however, its effects on precancerous lesions and the underlying mechanisms remain unclear. This study established a mouse model of precancerous pulmonary nodules and employed single-cell RNA sequencing (scRNA-seq) and immune repertoire sequencing (IR-seq) to elucidate the regulatory mechanisms by which smoking cessation influences the development of lung precancerous lesions.

MicroRNAs (miRNAs) regulate gene expression and generate isoforms (isomiRs) with distinct targeting properties, but their roles in cellular specification and reprogramming remain poorly understood. Here, we investigated miRNAs and isomiRs during direct reprogramming to conventional type 1 dendritic cells (cDC1s), identifying miR-124 and miR-142 as facilitators of the process. The canonical miR-124-3p strand induced a permissive chromatin landscape enriched for cooperative transcription factor motifs, thereby transiently increasing reprogramming efficiency. In contrast, miR-142 enhanced lineage fidelity by repressing fibroblast identity and activating cDC1-specific programs through cooperating isomiRs that drove XCR1 expression and increased type 3 interferon production. In addition, miR-142 mimics enhanced cancer cell reprogramming and anti-tumor immunity in vivo. Finally, we developed RNA-based cDC1 reprogramming using transcription factor mRNAs combined with miRNA mimics. Together, these findings uncover miRNA isoform diversity as a programmable regulatory layer in immune cell specification and establish RNA-driven reprogramming as a non-viral strategy for cancer immunotherapy.

Despite evidence highlighting immunoregulatory roles of tuft cells, it is not known whether vitamin A or retinoic acid affects tuft cell homeostasis. Here, we find that epithelial-intrinsic retinoic acid receptor regulates tuft cell numbers in mice. However, loss of retinoic acid signaling in tuft cells does not replicate this effect, suggesting altered differentiation. Consistent with this, retinoic acid administration to stem cell-derived organoids reduces tuft cell numbers, and epithelial-intrinsic retinoic acid is sufficient to alter tuft cell responses. In addition, dietary vitamin A dynamically alters tuft cells in vivo and regulates anti-helminth immunity in a tuft cell-dependent manner. Mechanistically, epithelial retinoic acid signaling promotes expression of Sprouty2, a negative regulator of tuft cell differentiation. Furthermore, stem cell-specific Sprouty2 disruption abrogates vitamin A regulation of tuft cells. Collectively, these data indicate that retinoic acid restricts tuft cell differentiation and carries important implications for how vitamin A affects intestinal health and immunity.

Nephrotoxicity is a major clinical concern because it can impair kidney function through inflammation and apoptosis. This study aimed to evaluate the effects of mesenchymal stem cell (MSC)-derived secretome on cyclooxygenase-2 (COX-2) and caspase-3 levels in a rat model of doxorubicin-induced nephrotoxicity.

Glioblastoma (GBM) is a highly lethal brain cancer in which the calcitonin receptor (CT Receptor), encoded by the CALCR gene, is expressed in 78-88% of patient biopsies. Here, we investigate whether the CT Receptor plays a role in cancer cell survival. In cancer cell lines, knockdown of CALCR disrupts the cell cycle and induces apoptosis, supporting an essential pro-survival role. The CALCR gene produces three main transcripts in humans, of which Transcript 1 encodes CALCRb mRNA including exon 10 and is translated into the CTb Receptor isoform, and Transcripts 2 and 3 which are translated into the CTa Receptor. CALCRb expression is conserved across a diverse range of mammalian species. We examined the expression of all CT Receptor isoforms (CALCRtotal) and CALCRb expression in four high-grade glioma stem-like cell lines and in U-87 MG glioblastoma cells. Using qPCR, we observed stable levels of both CALCRtotal and CALCRb expression under conditions of autophagy or apoptosis, consistent with a requirement for CALCRb in cell survival. As alternative splicing (AS) of key genes in cancers confers tumour resilience, we investigated AS of CALCR transcript 2 using long-read nanopore sequencing. Unexpectedly, we discovered a novel AS event causing inclusion of exon 10 within Transcript 2 in all glioblastoma cell lines investigated. This finding, together with stable CALCRb expression under cellular stress and the finding by other groups confirming that knockdown of CT Receptor compromises cell survival, implicates the CTb Receptor as a potential oncoprotein.

Primary non-response to anti-tumor necrosis factor (anti-TNF) therapy remains a major clinical challenge in ulcerative colitis (UC); however, public-data transcriptomic studies often yield unstable single-gene biomarkers and limited replication.

Preclinical evidence supports the potential use of human umbilical cord blood-derived mononuclear cells (hUCB-MNCs) for preterm infants with preterm birth-associated complications (PBAC). Allogeneic hUCB-MNCs could extend this therapeutic option to infants without autologous cord blood available at birth.

Myeloproliferative neoplasms (MPNs) are a group of hematologic malignancies for which current treatment options remain limited, underscoring the urgent need to explore novel therapeutic targets and intervention strategies. Through a high-throughput screen of an epigenetic compound library, we identified the SIRT6 allosteric agonist MDL-800 as a potent suppressor of neutrophil hyperplasia. We established an endogenous sirt6-mutant zebrafish model that develops a myeloproliferative neoplasm (MPN)-like phenotype, with a 64% incidence in adult zebrafish. Mechanistically, Sirt6 was found to regulate neutrophil proliferation in vivo and in vitro by deacetylating histone H3K9 at the c-myb promoter. Sirt6 deficiency led to aberrant proliferation of neutrophils and hematopoietic stem/progenitor cells, whereas Sirt6 overexpression significantly alleviated neutrophil hyperplasia and MPN-related symptoms. Furthermore, the SIRT6 activator MDL-800 enhanced the efficacy of imatinib and reduced neutrophil proliferation in a zebrafish leukemia model. In xenograft mouse models, the combination of MDL-800 and imatinib significantly inhibited leukemia progression and restored drug sensitivity in imatinib-resistant cases. This study establishes the Sirt6-c-Myb axis as a core epigenetic pathway for myeloid homeostasis, providing a novel strategy for simultaneously suppressing neutrophil hyperplasia and enhancing chemotherapeutic efficacy in hematologic malignancies.

Mesenchymal stem cells (MSCs) are multipotent, nonhematopoietic progenitors capable of supporting hematopoiesis and regulating immunity. Clinical studies have shown that MSCs can aid hematopoietic recovery and balance T helper 17 (Th17) and Treg cells in some aplastic anemia (AA) patients refractory to immunosuppressive therapy (IST). However, full clinical efficacy remains unachieved. To enhance MSC immune regulation while ensuring safety, previous research found interleukin-10 (IL-10), an immunosuppressive factor, can boost MSC function. In this study, we used lentiviral transduction to induce IL-10 gene overexpression in human umbilical cord-derived MSCs and validated their efficacy and safety in AA mouse models.

Atherosclerosis has traditionally been considered a lipid-driven disease. However, emerging evidence highlights the central role of inflammation in the development of atherosclerosis. Multiple cell types, including endothelial cells, macrophages, and other immune cells, interact within lesions to form a chronic inflammatory microenvironment. Unhealthy lifestyle, smoking and metabolic disorders like hyperlipidemia, hyperglycemia, and their derived pro-atherogenic products drive vascular inflammation. Recent evidence reveals that high-sensitivity C-reactive protein surpasses LDL-cholesterol in predicting future cardiovascular risk. Combining lipid-lowering with anti-inflammatory therapies significantly reduces event recurrence, underscoring the need for targeted drugs. Promising results have emerged from trials of anti-inflammatory agents. Statins and other lipid-lowering drugs also exhibit anti-inflammatory properties. However, cholesterol-independent strategies face challenges like infection risk from immunosuppression, requiring extensive safety evaluation. Enhancing intrinsic resilience mechanisms is a promising alternative in combating vascular inflammation and atherosclerosis. High-throughput screening accelerates drug development, while induced pluripotent stem cell-derived vascular organoids better simulate human atherosclerosis pathobiology, improving preclinical predictions. Research on organ interaction networks and trained immunity offers novel therapeutic targets. In this comprehensive review, we provide a state-of-the-art synthesis of the drivers, mechanisms, and potential therapies of atherosclerosis by targeting inflammation, with an aim to reducing residual cardiovascular risk in the post-statin era.

Acute myeloid leukemia (AML) is characterized by antigen heterogeneity and poor prognosis. Here, to tackle the heterogeneity of AML, we combined FLT3 with CD33 in a combinatorial OR-gate approach using our split, universal, programmable (SUPRA) chimeric antigen receptor (CAR) platform. The split platform affords tunability over activation levels and multiplexed targeting. We characterized the specificity and sensitivity of different SUPRA CAR adapters for each target across a panel of target cell lines. Our results demonstrate that this CAR system can effectively target two antigens with equivalent efficacy to conventional CARs while reducing the engineering burden of designing CAR T cells against multiple antigens. Furthermore, we can characterize an effective dose range where off-target cytotoxicity against hematopoietic stem and progenitor cells is minimized. Our SUPRA OR gate has the potential to provide an effective and safer solution to treating AML.

Renal ciliopathies encompass a spectrum of genetic disorders arising from structural or functional impairments of primary cilia, specialized organelles critical for mechanosensation and signal transduction within renal epithelial cells. These disorders are characterized by cystogenesis, driven by dysregulated ciliary signaling, leading to uncontrolled epithelial proliferation, aberrant growth, and loss of cellular polarity. The clinical trajectory evolves from initial cyst formation to advanced tubulointerstitial fibrosis and progressive renal failure. This progression is governed by pathogenic variants in genes encoding ciliary proteins. While advancements in genetic testing have established prenatal diagnosis as a pivotal tool for early identification, definitive diagnosis and therapeutic intervention remain challenging. These difficulties stem from several factors: incomplete understanding of the molecular mechanisms underlying cyst formation and fibrosis; limitations in prenatal diagnostic accuracy owing to phenotypic overlap and incomplete penetrance; and the marked genetic heterogeneity and diverse clinical trajectories of renal ciliopathies. Existing studies have primarily focused on unidirectional modulation of individual pathways, whereas the systematic integration of signaling network cascades remains largely unaddressed. This review systematically elucidates the molecular mechanisms and aberrant signaling pathways in renal ciliopathies, links genetic heterogeneity to clinical phenotypes, and lays a theoretical basis for prenatal diagnosis and novel therapies.

Tongue squamous cell carcinoma (TSCC) is a highly aggressive malignancy associated with unfavorable clinical outcomes, highlighting the critical need for dependable prognostic indicators. The epithelial-mesenchymal transition (EMT) is a fundamental biological process driving cancer progression. This study aimed to develop and independently validate an EMT-associated gene expression signature for predicting prognosis in TSCC through a multi-omics strategy.

Cryopreservation is essential for mesenchymal stem cell banking, but its effects on adipose-derived stem cell (ADSC) function remain debated. This study evaluated an optimised cryopreservation protocol for goat ADSCs using phenotypic, functional, mitochondrial-associated, conditioned-medium, proteomic, and wound-healing assays. Cryopreserved ADSCs retained morphology, viability, clonogenicity, proliferation, adherence, immunophenotype, and trilineage differentiation, comparable to those of fresh ADSCs. Post-thaw cells showed increased MitoTracker Green signal, JC-1 red/green ratio, ATP content, relative mtDNA abundance, and SOD2-associated immunofluorescence, together with lower H2O2-induced ROS- and autophagy-associated fluorescence. Conditioned-medium proteomics suggested exploratory shifts toward extracellular matrix-, anti-protease-, and redox-associated proteins. Functionally, cryopreserved ADSCs maintained inflammatory cue-directed migration, modulated macrophage surface marker expression, and reduced oxidative stress-associated injury readouts in goat dermal fibroblasts. In a goat excisional wound model, ADSC treatment improved wound-closure kinetics and supported histological repair. Overall, optimised cryopreservation preserved the functional competence of goat ADSCs and was associated with stress-adapted post-thaw features.

Thyroid hormone (TH) signaling is widely recognized as an important regulator of skeletal muscle regeneration, yet the mechanisms by which aging alters TH-dependent regenerative responses remain unresolved. Current models generally assume that age-associated regenerative decline reflects reduced thyroid hormone receptor (THR) signaling resulting from alterations in hormone availability, receptor expression, or downstream transcriptional activity. However, several observations challenge this view, including the persistence of THR activity in aged tissues, the limited efficacy of thyroid hormone replacement in restoring regeneration, and the distinct regenerative phenotypes observed in aging and hypothyroidism. Here, we propose the hypothesis that aging does not primarily impair skeletal muscle regeneration through loss of THR signaling, but through disruption of endocrine-immune-niche integration. In this framework, THR signaling functions as a systems-level coordinator that integrates endocrine signals with stromal remodeling, immune responses, and intercellular communication networks required for effective tissue repair. To evaluate whether available evidence is consistent with this hypothesis, we integrate published studies with analyses of publicly available murine single-cell transcriptomic, chromatin-accessibility, and ligand-receptor datasets spanning skeletal muscle regeneration, aging, and hypothyroidism. These associative analyses suggest that substantial components of THR activity remain detectable during aging, whereas coordination between THR signaling, macrophage remodeling, extracellular-matrix programs, inflammatory pathways, and Notch-mediated communication is altered. The findings are associative and do not establish causality; rather, they provide a hypothesis-generating framework for experimental testing. We propose that aging represents a state of impaired endocrine-immune-niche integration in which thyroid hormone signaling remains partially active but becomes increasingly uncoupled from the regenerative programs it normally coordinates. This framework offers a possible unifying explanation for several previously disconnected observations and generates experimentally testable predictions regarding endocrine-immune communication during tissue repair and aging.

Volumetric muscle loss (VML) causes irreversible loss of contractile tissue and creates a hostile regenerative niche marked by sustained inflammation, oxidative stress, fibrosis, and poor functional recovery. Here, we develop an injectable and photocurable microsphere-in-hydrogel platform that couples structural support with sustained small-molecule immunoregulation. To counteract this hostile microenvironment, we utilized emodin, a natural anthraquinone recognized for its potent anti-inflammatory and reactive-oxygen-species-scavenging properties. Emodin-loaded sodium alginate microspheres were generated via ionic crosslinking and embedded within a gelatin methacryloyl matrix to form E-AMs@GM. The composite hydrogel exhibited defect-conforming moldability, porous microarchitecture, tunable swelling/degradation, and broad interfacial adhesion. In vitro, E-AMs@GM showed excellent cytocompatibility and attenuated intracellular reactive oxygen species in human bone-marrow-derived mesenchymal stem cells under oxidative challenge. In macrophages, E-AMs@GM reduced pro-inflammatory activation while enhancing pro-regenerative programs, accompanied by decreased inflammatory cytokines and increased interleukin-10. E-AMs@GM also promoted C2C12 myogenic differentiation and myotube maturation. In a murine VML model, E-AMs@GM alleviated inflammation and fibrotic remodeling, increased myogenic progenitor activity and myofiber regeneration, and improved locomotor performance by CatWalk analysis. Mechanistically, scRNA sequencing revealed that E-AMs@GM enriches a reparative Mmp12+ macrophage subset and rewires macrophage-muscle satellite cell interactions through SPP1-CD44 and SPP1-integrin signaling, consistent with accelerated transition from inflammatory clearance to tissue reconstruction. Together, this depot-enabled hydrogel provides an instructive biomaterial strategy for functional VML repair.